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Fecal contamination source tracking in sediment using antibiotic resistance patterns

Background:. EPA standards ? 500CFU maxPer 100 ml sampleJesters Creek exceeds standardsSource determination not easyMethodology needed. Antibiotic Resistance Patterns. Antibiotics are everywhere!Sources of antibioticsNature (bacteria, molds, etc.)VeterinariansDoctorsBacteria are everywhere

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Fecal contamination source tracking in sediment using antibiotic resistance patterns

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    1. Fecal contamination source tracking in sediment using antibiotic resistance patterns Margaret Grace Mills Clayton College and State University Summer 2004

    2. Background: EPA standards ? 500CFU max Per 100 ml sample Jesters Creek exceeds standards Source determination not easy Methodology needed

    3. Antibiotic Resistance Patterns Antibiotics are everywhere! Sources of antibiotics Nature (bacteria, molds, etc.) Veterinarians Doctors Bacteria are everywhere! Antibiotic resistant strains Conjugate and pass on resistance Resistance from mutations (with selection factor present) Some bacteria are naturally resistant (species specific – genetics of organism) G+ vs. G-G+ vs. G-

    4. How can we use this? Bacterial resistance not the same in every organism Use statistics to compare antibiotic resistance patterns

    5. Why do we care? Fecal coliform numbers increase dramatically after heavy rain Cause? Run-off water contamination Sediment contamination Method – use sediment samples to try and isolate coliforms Too many words Nutrients in the sediment – more bacteriaToo many words Nutrients in the sediment – more bacteria

    6. Sample Collecting Pasteur pipette

    7. Sediment Sampling

    8. Antibiotic plates Collected 201 isolates – frozen stock cultures made Eleven antibiotics in three concentrations 34 characteristics Each also had a control plate There were 70 plates per micro well plate! Characteristics made a ARPCharacteristics made a ARP

    9. My plates

    10. Replication A 48-prong replicator was used to transfer bacteria antibiotic plates.

    11. Growth vs. Non-growth Put this on slide 11Put this on slide 11

    12. Scoring Plates A spreadsheet was made with the growth data for statistical analysis (see next slide).

    14. Database A database was made using fecal coliforms isolated from fecal samples Cases classified correctly 52% sewage 71% wild 61% horse 70% dog

    15. Antibiotics chosen for DFA Oxytetracycline 25mg/ml Oxytetracycline 200mg/ml Ampicillin 0.3mg/ml Doxycycline 64mg/ml Streptomycin 20mg/ml Penicillin G 20mg/ml Cephalothin 128mg/ml

    16. Classification Results (with chosen antibiotics) Make chart of only my data – not MyHangsMake chart of only my data – not MyHangs

    17. Discussion Similar to last year Leaky sewer? Run-off? Silt fence not adequate to stop contamination Three purple isolates Possible new strain Two classified as sewage, one as dog Humans classified poorly – can lead to increase of cases classified as “human”Humans classified poorly – can lead to increase of cases classified as “human”

    18. Discussion

    19. Acknowledgements Dr. Michelle Furlong Clayton College and State University Dr. Stephen Burnett Dr. Valerie Harwood Dave Stuchkus Clayton County Water Authority Kim Zimmerman

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